LOCUS       Exported                4757 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses thermostable variant of human carbonic anhydrase II with 
            TAG stop codon at site 93 and C-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-CA TAG93
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  TITLE     Bacterial expression plasmids for genetic code expansion laboratory 
            course
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4757
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        11..295
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     218..237
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1109..1126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1168..1185)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1338..1424
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1516..1543
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1562..1653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1654..2514
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1872..1891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2685..3273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3174..3193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3427..3444
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3459..3599
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3525..3544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3685..3707
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3863..4741)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
       61 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
      121 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
      181 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
      241 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
      301 ttttttgggc taacaggagg aattaaccat ggcgcatcat tggggttacg gtaaacacaa
      361 cggtccggag cattggcaca aagattttcc aattgcgaag ggcgaacgtc aaagcccggt
      421 tgacattgat acgcacacgg caaagtacga cccgagcctg aaaccgctga gcgtttccta
      481 tgaccaggct acgagcctgc gtatcctgaa caatggccac accttcaacg tggagtttga
      541 tgattcccaa gataaggcgg ttctgaaagg tggtccgttg gatggcacct accgcctgat
      601 ccaatagcac tttcactggg gtagccacga cggtcagggc agcgagcata ccgtggacaa
      661 aaagaagtat gcagccgaac tgcacctggt gcattggaac acgaagtacg gcgacttcgg
      721 taaagcggtc cagcaaccgg acggtctggc tgttctgggt attttcctga aggtcggcag
      781 cgcgaacccg ggtctgcaga aagtggttga cgtgttggac tctatcaaga ccaaaggcaa
      841 gagcgcggac ttcaccaatt tcgatccgcg tggtctgctg ccggagagcc tggattactg
      901 gacttatccg ggcagcctga ccaccccgcc attgctggag tgcgtgacct ggatcgtctt
      961 gaaagaaccg atcagcgtta gctctgaaca ggtcagcaag ttccgcaagc tgaatttcaa
     1021 tggtgagggc gagccggaag aaccgatggt cgataattgg cgtcctaccc aaccgctgaa
     1081 aaaccgccag attaaagcat cctttaagca tcaccaccac catcactaaa agcttggctg
     1141 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     1201 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     1261 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     1321 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1381 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     1441 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     1501 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     1561 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1861 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     1921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2341 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2821 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3301 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3361 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3421 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     3481 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     3541 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     3601 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3661 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3721 caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat
     3781 gtgcctgtca aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt
     3841 caattgtctg attcgttacc aattatgaca acttgacggc tacatcattc actttttctt
     3901 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     3961 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     4021 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     4081 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     4141 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     4201 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     4261 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     4321 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     4381 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     4441 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     4501 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     4561 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     4621 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     4681 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     4741 tacttttcat actcccg
//
